LOCUS       Exported                8504 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGBT-PX, complete sequence.
ACCESSION   HQ335166
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8504)
  AUTHORS   Clark KJ, Balciunas D, Pagoda H-M., Ding Y, Westcot SE, Bedell VM, 
            Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielson A, 
            Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC.
  TITLE     In vivo protein trapping produces a functional expression codex of 
            the vertebrate proteome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8504)
  AUTHORS   Clark KJ, Balciunas D, Pagoda H-M., Ding Y, Westcot SE, Bedell VM, 
            Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielson A, 
            Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-OCT-2010) Biochemistry and Molecular Biology, Mayo 
            Clinic, 221 4th Ave. SW, Rochester, MN 55902, USA
REFERENCE   3  (bases 1 to 8504)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8504
                     /organism="Cloning vector pGBT-PX"
                     /mol_type="other DNA"
                     /db_xref="taxon:1001535"
     mobile_element  complement(1..192)
                     /mobile_element_type="transposon:Tol2"
                     /note="mini ITR-R"
     misc_recomb     227..260
                     /label=loxP
                     /note="loxP"
     protein_bind    227..260
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          313..410
                     /note="carp beta-actin intron 1 splice acceptor"
     regulatory      504..1769
                     /regulatory_class="polyA_signal_sequence"
                     /note="ocean pout antifreeze poly(A) signal; terminator; 
                     putative border element"
     rep_origin      1588..2044
                     /label=F1 Ori
                     /note="F1 Ori"
     promoter        1741..1759
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      1768..2903
                     /regulatory_class="promoter"
                     /note="carp beta-actin promoter region"
     exon            2909..3000
                     /note="carp beta-actin exon 1 (non-coding)"
     intron          3001..4264
                     /note="carp beta-actin intron 1 splice acceptor"
     CDS             4329..5045
                     /codon_start=1
                     /product="green fluorescent protein GM2 variant"
                     /label=green fluorescent protein GM2 variant
                     /note="GFP"
                     /protein_id="AEA29850.1"
                     /translation="MSKGEELFTGVVPILVELNGDVNGHKFPVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     intron          5100..5236
                     /note="carp beta-actin intron 1 splice donor"
     protein_bind    5254..5287
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     mobile_element  5304..5565
                     /mobile_element_type="transposon:Tol2"
                     /note="mini ITR-L"
     promoter        complement(5578..5596)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5603..5619)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5760..6215
                     /label=F1 Ori
                     /note="F1 Ori"
     promoter        6293..6397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6398..7258
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="bla"
                     /protein_id="AEA29849.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            6398..7258
                     /gene="ampR"
                     /label=ampR
     rep_origin      complement(7319..8199)
                     /direction=LEFT
                     /label=ColE1 Ori
                     /note="ColE1 Ori"
     rep_origin      7429..8017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8305..8326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8341..8371
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8379..8395
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8403..8419
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8437..8455
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"